<i>Gilgyung-tang</i> Inhibits the Migration and Invasion of Human Bladder Cancer 5637 Cells through the Tightening of Tight Junctions and Inhibition of Matrix Metalloproteinase Activity

Hong, Su-hyun; Choi, Yung-hyun

The Journal of Internal Korean Medicine > Volume 37(1); 2016 > Article

Original Article

The Journal of Internal Korean Medicine 2016;37(1): 16-25.

길경탕의 치밀결합 강화 및 MMPs의 활성 억제를 통한 인체방광암세포의 이동성 및 침윤성의 억제

홍수현¹, 최영현^1,2

¹동의대학교 한의과대학 생화학교실
²동의대학교 항노화연구소 및 블루바이오소재개발센터

Gilgyung-tang Inhibits the Migration and Invasion of Human Bladder Cancer 5637 Cells through the Tightening of Tight Junctions and Inhibition of Matrix Metalloproteinase Activity

Su-hyun Hong¹, Yung-hyun Choi^1,2

¹Dept. of Biochemistry, College of Korean Medicine, Dong-Eui University
²Anti-Aging Research Center & Blue-Bio Industry RIC, Dong-Eui University

Correspondence

Yung-hyun Choi ,Tel: 051-850-7413 , Fax: 051-853-4036, Email: choiyh@deu.ac.kr

Received: January 8, 2016, Accepted: April 1, 2016, Published online: March 31, 2016.

ABSTRACT

Objectives:
Gilgyung-tang (GGT) has been used as one of the main multi-herb formulas to treat “Peo-ong” (lung abscess). In this study, we investigated the inhibitory effects of water extracts of GGT on cell migration and invasion, two critical cellular processes that are often deregulated during metastasis, in human bladder cancer 5637 cells.

Methods:
Effects on cell viability were quantified using an MTT assay. To analyze the anti-metastatic effects, we conducted a wound healing migration assay, an in vitro invasiveness assay, and a measurement of the transepithelial electrical resistance (TER). The expression of protein and mRNA were measured by Western blotting and real-time polymerase chain reaction (RT-PCR), respectively.

Results:
GGT markedly inhibited the cell motility and invasiveness of 5637 cells within the concentration range that was not cytotoxic. The inhibitory effects of GGT on cell invasiveness were associated with tightening of the tight junctions (TJs), which was demonstrated by an increase in the TER. The RT-PCR and Western blotting results indicated that GGT decreased the levels of claudin proteins. GGT also inhibited the activity and expression of matrix metalloproteinase (MMP)-2 and -9 and simultaneously increased the levels of tissue inhibitor of metalloproteinase-1 and -2.

Conclusions:
Our findings suggest that GGT reduces both the migration and the invasion of 5637 cells by modulating the activity of TJs and MMPs.

Key words: Gilgyung-tang, invasion, tight junctions, claudin, matrix metalloproteinase